长链非编码RNA OSER1-AS1对缺氧环境下胃癌细胞上皮间质转化的机制研究

Mechanism of LncRNA OSER1-AS1 on epithelial-mesenchymal transition ofgastric cancer cell induced by Hypoxia

目的:探讨长链非编码RNA氧化应激反应丝氨酸丰富1反义RNA 1(LncRNA OSER1-AS1)对缺氧诱导的胃癌细胞上皮间质转化(EMT)的作用机制。方法:缺氧组(Hypoxia)、对照组(Hypoxia+NC)细胞转染si-LncRNA OSER1-AS1-NC,实验组(Hypoxia+si)细胞转染si-LncRNA OSER1-AS1后,Hypoxia、Hypoxia+NC、Hypoxia+si组细胞的培养基中添加CoCl2构建缺氧微环境;另取细胞转染si-LncRNA OSER1-AS1-NC后,常规培养,为正常组(control)细胞。qRT-PCR检测各组细胞中LncRNA OSER1-AS1、E-cadherin和α-SMA mRNA表达,检测各组细胞形态的改变,CCK-8法检测细胞的增殖能力;Transwell侵袭检测各组细胞的侵袭能力;免疫荧光和Western blot检测各组细胞中E-cadherin、α-SMA以及对应蛋白的表达水平。结果:与control组相比,Hypoxia、Hypoxia+NC、Hypoxia+si组细胞中LncRNA OSER1-AS1的表达、E-cadherin的mRNA和蛋白的表达以及细胞中E-cadherin与α-SMA荧光强度的比值明显降低,细胞中发生EMT细胞比例、细胞增殖性能、侵袭能力、α-SMA的mRNA和蛋白表达明显升高,与Hypoxia+NC组相比,Hypoxia+si组细胞中LncRNA OSER1-AS1的表达、E-cadherin的mRNA和蛋白表达以及细胞中E-cadherin与α-SMA荧光强度的比值明显降低,细胞中发生EMT细胞比例、细胞增殖性能、侵袭能力、α-SMA的mRNA和蛋白表达明显升高,差异均具有统计学意义(均P<0.05)。结论:LncRNA OSER1-AS1在胃癌组织中呈低表达,沉默其表达RNA能明显增强其侵袭能力,推动肿瘤细胞的EMT过程。

Objective:To investigate the mechanism of lncRNA oxidative stress responsive serine rich 1 antisense(LncRNA OSER1-AS1)on epithelial-mesenchymal transition(EMT)of gastric cancer cell induced by Hypoxia.Methods:Hypoxia group(Hypoxia),control group(Hypoxia+NC)cells were transfected with si-LncRNA OSER1-AS1-NC,experimental group(Hypoxia+si)cells were transfected with si-LncRNA OSER1-AS1.After transfection,CoCl2 was added to the culture medium of Hypoxia,Hypoxia+NC,Hypoxia+si group to build a hypoxic microenvironment.Another cell was transfected with si-LncRNA OSER1-AS1-NC and cultured routinely to be control cells.The mRNA expression of LncRNA OSER1-AS1,E-cadherin andα-SMA in each group were detected by qRT-PCR,and the changes of cell morphology were detected.The proliferation ability of cells was detected by CCK-8 method.The invasion ability of each group was detected by Transwell invasion.The expression levels of E-cadherin,α-SMA and corresponding proteins were detected by immunofluorescence and Western blot.Results:Compared with the control group,LncRNA OSER1-AS1 expression,E-cadherin mRNA and protein expression and the ratio of E-cadherin toα-SMA fluorescence intensity in Hypoxia,Hypoxia+NC and Hypoxia+si groups were significantly decreased.The proportion of EMT cells,cell proliferation performance,invasion ability,α-SMA mRNA and protein expression were significantly increased in the cells,and compared with the Hypoxia+NC group,The expression of LncRNA OSER1-AS1,mRNA and protein expression of E-cadherin and the ratio of E-cadherin toα-SMA fluorescence intensity in the Hypoxia+si group were significantly decreased.The proportion of EMT cells,cell proliferation ability,invasion ability,α-SMA mRNA and protein expression were significantly increased,and the differences were statistically significant(all P<0.05).Con⁃clusion:LncRNA OSER1-AS1 is under-expressed in gastric cancer tissues,and silencing its expression can significantly enhance the invasion ability and promote the EMT process of tumor cells.